Li Bo-lun, Hou Jun-jie, Nie Fang-fei, Qin Ze-lian, Zhao Xia, Zhang Zhe, Ma Yong-guang
Department of Plastic Surgery, Peking University Third Hospital, Beijing 100191, China. Corresponding author: Qin Ze-lian, Email:
Zhonghua Wei Zhong Bing Ji Jiu Yi Xue. 2013 Sep;25(9):523-6. doi: 10.3760/cma.j.issn.2095-4352.2013.09.004.
To investigate the expressions of periostin (PN), angiopoietin-1 (Ang-1), vascular epithelial growth factor (VEGF) and fetal liver kinase-1 (Flk-1) during the processes of scar formation and modulation in rat cutaneous wounds and probe into their roles in wound healing and scaring.
Eighty-two male Sprague-Dawley (SD) rats were randomly divided into 10 groups with 8-9 rats in each group. Two 2 cm×2 cm full-thickness excisional wounds in the back were created in each rat. The wound surface was observed, and the healing area was measured. The pathological change was observed after hematoxylin and eosin (HE) staining. The expressions of PN, Ang-1, VEGF and Flk-1 in wound surface scar at 4-8 weeks were determined with immunohistochemistry. The expressions of PN, Ang-1 and VEGF were determined by Western blotting. The normal skin was served as control.
HE staining showed that the wound surface tissue had healed with epithelization at 4-8 weeks. Immunohistochemistry results showed that there was no significant difference in Flk-1 expression between wound surface tissue and normal skin. The PN expression (A value/μm(2)) in wound surface tissue was significantly lower than that in normal skin at 5 weeks (2.43±0.44 vs. 4.24±0.50, P<0.05), and the expression of Ang-1 and VEGF (A value/μm(2)) at 4, 5, 6, 8 weeks was significantly lower than that in normal skin (Ang-1: 3.51±0.93, 3.10±0.57, 2.77±0.59, 2.77±1.26 vs. 4.89±0.48; VEGF: 1.76±0.68, 1.75±0.49, 1.99±0.42, 1.94±0.86 vs. 4.86±1.63, all P<0.05). In wound surface scar, PN and Flk-1 positive signal was found in cell, and the Ang-1 and VEGF positive signal in extracellular matrix. Western blotting data demonstrated that the expressions of PN, Ang-1 and VEGF peaked at the 10th day after excision with increases to 7.90-22.56 folds compared with normal skin (PN: 2.45±1.51 vs. 0.31±0.19, Ang-1: 18.43±15.20 vs. 1.53±1.42, VEGF: 6.09±4.66 vs. 0.27±0.13, P<0.05 or P<0.01), and then followed with a decrease.
PN, Ang-1, VEGF and Flk-1 are transiently overexpressed in early stage of full-thickness cutaneous wound healing in rats. Their expressions vary in wounds and scars. They participate in the healing of full-thickness cutaneous wounds together and may be essential for the proliferation stage during wound healing.
研究骨膜蛋白(PN)、血管生成素-1(Ang-1)、血管内皮生长因子(VEGF)和胎儿肝激酶-1(Flk-1)在大鼠皮肤伤口瘢痕形成及重塑过程中的表达情况,并探讨它们在伤口愈合和瘢痕形成中的作用。
82只雄性Sprague-Dawley(SD)大鼠随机分为10组,每组8 - 9只。在每只大鼠背部制作两个2 cm×2 cm的全层切除伤口。观察伤口表面情况,测量愈合面积。苏木精-伊红(HE)染色后观察病理变化。采用免疫组织化学法检测4 - 8周时伤口表面瘢痕中PN、Ang-1、VEGF和Flk-1的表达。采用蛋白质免疫印迹法检测PN、Ang-1和VEGF的表达。以正常皮肤作为对照。
HE染色显示,4 - 8周时伤口表面组织已上皮化愈合。免疫组织化学结果显示,伤口表面组织与正常皮肤中Flk-1表达无显著差异。伤口表面组织中PN表达(A值/μm²)在第5周时显著低于正常皮肤(2.43±0.44 vs. 4.24±0.50,P<0.05),Ang-1和VEGF表达(A值/μm²)在第4、5、6、8周时显著低于正常皮肤(Ang-1:3.51±0.93,3.10±0.57,2.77±0.59,2.77±1.26 vs. 4.89±0.48;VEGF:1.76±0.68,1.75±0.49,1.99±0.42,1.94±0.86 vs. 4.86±1.63,均P<0.05)。在伤口表面瘢痕中,PN和Flk-1阳性信号见于细胞内,Ang-1和VEGF阳性信号见于细胞外基质。蛋白质免疫印迹数据显示,PN、Ang-1和VEGF表达在切除后第10天达到峰值,与正常皮肤相比增加至7.90 - 22.56倍(PN:2.45±1.51 vs. 0.31±0.19,Ang-1:18.43±15.20 vs. 1.53±1.42,VEGF:6.09±4.66 vs. 0.27±0.13,P<0.05或P<0.01),随后下降。
PN、Ang-1、VEGF和Flk-1在大鼠全层皮肤伤口愈合早期短暂过表达。它们在伤口和瘢痕中的表达有所不同。它们共同参与全层皮肤伤口的愈合,可能对伤口愈合的增殖阶段至关重要。